Method and device for manufacturing tissue section

ABSTRACT

The method for manufacturing a tissue section, which is used for a sample for observation by a microscope, comprises the steps of slicing an organism specimen ( 12 ), whose form has been fixed by freezing or by using an embedding agent, along a slicing surface, characterized by: adjusting a distance between the slicing surface of the organism specimen ( 12 ) and one side of a tape ( 32 ) and a temperature difference between the organism specimen ( 12 ) and the tape ( 32 ) when the slicing of the organism specimen ( 12 ) is started, thereby a tip part of the tissue section ( 16 ) curling to the outside of the slicing surface of the organism specimen ( 12 ) is allowed to adhesively abut on one side of the tape ( 32 ) running apart from the slicing surface of the organism specimen ( 12 ); and running the tape ( 32 ) at a speed in synchronism with a slicing speed of the tissue section ( 16 ) after the tip part of the tissue section ( 16 ) is allowed to adhesively abut on the one side of the tape ( 32 ), thereby the entire tissue section ( 16 ) cut off from the organism specimen ( 12 ) can be adhered onto the one side of the tape ( 32 ).

FIELD OF TECHNOLOGY

The present invention relates to a method and a device for manufacturinga tissue section, more precisely relates to a method and a device formanufacturing a tissue section, which is used for a sample forobservation by a microscope and made by slicing an organism specimen,whose form has been fixed by freezing or by using an embedding agent,along a slicing surface.

BACKGROUND TECHNOLOGY

Pathological samples are observed by the steps of: slicing an organismspecimen to form tissue sections, adhering the tissue sections ontoslide glasses to form the pathological samples; and observing thesamples by a microscope.

To observe by the microscope, the tissue sections must be thin so as toallow lights to pass through them. Thus, a form of the organismspecimen, from which the tissue sections are cut, has been previouslyfixed by freezing or by using an embedding agent, e.g., paraffin, so asto easily slice the organism specimen.

However, even if the form of the organism specimen is fixed by freezingor by using the embedding agent as shown in FIG. 5, the tissue section16 (upwardly) curls to the outside of a slicing surface of the organismspecimen 12 when the organism specimen 12, which is mounted on a table12 and whose form is fixed, is moved in a direction of an arrow and cutby a fixed knife 10.

When the curling tissue section 16 is mounted on a flat slide glass,creases are formed, a bad sample for microscope observation is oftenproduced.

Especially, degree of curling of the tissue section 16, which is made byslicing the organism specimen 12 whose form has been fixed by freezing(hereinafter sometimes referred to “frozen organism specimen 12”) issometimes greater than that of the tissue section 16, which is made byslicing the organism specimen 12 whose form has been fixed by theembedding agent, e.g., paraffin, (hereinafter sometimes referred to“embedded organism specimen 12”).

The tissue sections 16 for an urgent pathological examination is usuallycut from the frozen organism specimen 12 because there is no time to fixthe form by the embedding agent, e.g., paraffin.

Conventionally, as shown in FIG. 6A, the table 14, on which the frozenorganism specimen 12 is mounted, is moved in the direction of the arrow,and the tissue section 16, which has been cut by the fixed knife 10, ismanually adhered to a tip part of a brush 100, then the brush 100holding the tissue section 16 at the tip part is synchronously movedwith a slicing speed, as shown in FIG. 6B, so as to restrict the curl ofthe tissue section 16.

To manually perform the steps shown in FIGS. 6A and 6B, enough skill isrequired, and it is often impossible to reobtain the same frozenorganism specimen 12, from which the tissue section 16 will be cut.Therefore, miscutting the tissue section 16 is not allowed, so a workermust cut carefully. Further, there is a risk of infection from thefrozen organism specimen 12.

To execute a rapid and correct diagnosis on the basis of a rapidexamination of the tissue section 16 cut from the frozen organismspecimen 12, experienced pathological doctors are required, sohospitals, which are capable of rapidly examining the frozen organismspecimens 12, are limited.

In the case of treating the embedded organism specimen 12 too, it isdifficult for a nonskilled worker to manually perform the steps shown inFIGS. 6A and 6B, it is often impossible to reobtain the same embeddedorganism specimen 12, from which the tissue section 16 will be cut, andmiscutting the tissue section 16 is not allowed, so a worker must cutcarefully as well as the frozen organism specimen 12.

To solve the disadvantages, some methods for manufacturing tissuesections from the conventional organism specimens 12 are disclosed in,for example, Japanese Patent Gazettes NO. 4-177143, No. 7-159298 and No.2002-31586, in each of which an adhesive face of a transparent film isadhered on the organism specimen, the organism specimen is slicedimmediately under the transparent film by, for example, a knife, thenthe tissue section is taken out together with the transparent film. Inthe patent gazettes No. 4-177143 and No. 7-159298, the transparent filmis pressed onto the organism specimen by a roller or a plunger when thetransparent film is adhered onto the organism specimen.

Further, in a Japanese Patent Gazette No. 6-323967, a tissue section,which has been cut from an organism specimen, is dipped into water, thenthe tissue section floating on a water surface is scooped up by atransparent film.

By employing the methods disclosed in the patent gazettes, the tissuesection can be adhered onto one side of the transparent film withoutmanually performing the difficult steps shown in FIGS. 6A and 6B.

However, since the adhesive face of the transparent film is pressed ontothe organism specimen, an adhesive directly contacts the organismspecimen so that tissue cells, which are in and near the contact surfaceof the organism specimen, are sometimes badly influenced andtransformed.

Further, if air is left in a space between the transparent film and theorganism specimen, creases are easily formed in the tissue section cutoff from the organism specimen, so that it is necessary to discharge theair by pressing the transparent film onto the organism specimen with aprescribed force. Tissue cells of the organism specimen are easilybroken by pressing the transparent film, and thickness of the tissuesection, which is cut off from the organism specimen, is partiallyvaried by repeatedly pressing and releasing the organism specimen.

When a plurality of tissue sections are sliced off from one organismspecimen to manufacture a plurality of microscopic samples, transparentfilms are pressed onto the organism specimen for each slice so as todischarge air. Further, the tissue sections must be adhered on one sideof the transparent film in slicing order so as to individually recognizethe tissue sections.

Therefore, the steps until adhering the tissue sections on the one sideof the transparent film in the slicing order are included in arate-controlling stage of manufacturing microscopic samples.

In the method such that the tissue section is scooped from water by thetransparent film, frozen organism specimens cannot be used, so themethod is limited to organism specimens embedded by a hydrophobicembedding agent, e.g., paraffin.

Further, in the case of using the embedded organism specimens embeddedby the hydrophobic embedding agent, e.g., paraffin, if a plurality oftissue sections are dipped into water, it is difficult to individuallyrecognize them, so one tissue section is dipped into the water after theprior tissue section is scooped by the transparent film. Therefore, thestep of making the tissue sections is the rate-controlling stage ofmanufacturing microscopic samples.

An object of the present invention is to provide a method and a devicefor manufacturing a tissue section, which are capable of treating afrozen organism specimen and an embedded organism specimen and rapidlyadhering the tissue section onto a film without pressing the film.

DISCLOSURE OF THE INVENTION

The inventors of the present invention have studied and found that a tippart of a tissue section curls to outside of a slicing surface of anorganism specimen when slicing the organism specimen is started.Further, they found that the tissue section can be rapidly adhered ontoa film without pressing the tissue section onto the film by adhesivelyabutting the tip part of the tissue section on one side of the filmrunning apart from the organism specimen and running the film at a speedin synchronism with a slicing speed of the tissue section, so that theyhave invented the present invention.

Namely, the method for manufacturing a tissue section, which is used fora sample for observation by a microscope, comprises the steps of slicingan organism specimen, whose form has been fixed by freezing or by usingan embedding agent, along a slicing surface, is characterized by:adjusting a distance between the slicing surface of the organismspecimen and one side of a film and a temperature difference between theorganism specimen and the film when the slicing of the organism specimenis started, thereby a tip part of the tissue section curling to theoutside of the slicing surface of the organism specimen is allowed toadhesively abut on one side of the film running apart from the slicingsurface of the organism specimen; and running the film at a speed insynchronism with a slicing speed of the tissue section after the tippart of the tissue section is allowed to adhesively abut on the one sideof the film, thereby the entire tissue section cut off from the organismspecimen can be adhered onto the one side of the film.

And, the device for manufacturing a tissue section, which is used for asample for observation by a microscope, comprising: means for slicing anorganism specimen, whose form has been fixed by freezing or by using anembedding agent, along a slicing surface; and means for running a filmapart from the slicing surface of the organism specimen, ischaracterized by: means for adjusting a distance between the slicingsurface of the organism specimen and one side of the film so as to makea tip part of the curling tissue section contact with the outside of theslicing surface of the organism specimen when the slicing of theorganism specimen is started; means for adjusting temperature betweenthe organism specimen and the film so as to make the tip part of thetissue section, which has been contacted the outside of the film,adhesively abut on the film; and means for synchronizing a slicing speedof the tissue section, whose tip part has been adhered on the one sideof the film, and a running speed of the film so as to adhere the entiretissue section cut off from the organism specimen onto the one side ofthe film.

In the present invention, the temperature difference between theorganism specimen and the film may be adjusted by adjusting temperatureof the film and/or temperature of an atmosphere, in which the organismspecimen is sliced, so that the temperature difference between theorganism specimen and the film can be easily adjusted. Preferably,temperature of the film is adjusted to temperature, at which a part ofan ice or the embedding agent for fixing the form of the organismspecimen melts and the tissue section is adhered onto the film.

Temperature of an atmosphere, in which the organism specimen is sliced,may be adjusted so as to hold the fixed form of the organism specimen,so that slicing the organism specimen can be performed stably.

A plurality of rollers including a close roller, which is providednearest to the slicing surface of the organism specimen, may be providedin a film running path, and the distance between the slicing surface ofthe organism specimen and the one side of the film may be adjusted bymoving the close roller to and away from the slicing surface of theorganism specimen, so that the distance between the slicing surface ofthe organism specimen and the one side of the film can be easilyadjusted.

If a center of the close roller is moved on a bisector line of anintersection angle between a line extended from a cutting surface of acutting tool, which intersects a line extended from the slicing surfaceof the organism specimen, and the line extended from the slicing surfaceof the organism specimen, the close roller never interferers withcutting means and the slicing surface of the organism specimen.

Further, the film running speed with respect to the slicing speed of thetissue section is adjusted so as not to form creases in the tissuesection and break the tissue section, namely, preferable ratio (Vt/Vs)of the film running speed (Vt) to the slicing speed (Vs) of the tissuesection is 1.2-0.8.

Note that, if the film is a transparent film, the tissue section adheredon the transparent film can be used as a microscopic sample.

In the present invention, since the tip part of the tissue section curlsto outside of the slicing surface of the organism specimen when theslicing of the organism specimen is started, so that the entire tissuesection cut off from the organism specimen can be adhered onto the oneside of the film running apart from the slicing surface of the organismspecimen.

Namely, when the slicing of the organism specimen is started, the tippart of the tissue section curling to the outside of the slicing surfaceof the organism specimen abuts on the one side of the film running apartfrom the organism specimen. The temperature difference between theorganism specimen and the film has been adjusted so as to adhere the tippart of the tissue section on the one side of the film.

Therefore, the tissue section, whose tip part has been adhered on theone side of the film, can be entirely adhered onto the one side of thefilm by running the film at the speed in synchronism with the slicingspeed of the tissue section.

As described above, since the entire tissue section can be rapidlyadhered on the film running apart from the organism specimen, the filmis not pressed onto the organism specimen, so that no tissue cells ofthe organism specimen are broken and the tissue section having uniformthickness can be stably manufactured.

Further, since the adhesion of the tissue section is controlled on thebasis of the temperature difference between the film and the organismspecimen, no adhesive is applied to the one side of the film so that notissue cells are badly influenced and transformed by adhesive. In thepresent invention, the tissue section is immediately adhered onto thefilm after the tissue section is cut off from the organism specimen;even if a plurality of the tissue sections are sliced, the tissuesections can be adhered onto the film in slicing order, so that thetissue section can be easily individually recognized.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic view of a tissue section manufacturing device ofan embodiment of the present invention; FIG. 2 is an explanation viewexplaining a method of adjusting a distance C between a close roller 38of the device shown in FIG. 1 and a slicing surface of an organismspecimen 12; FIGS. 3A-3E are explanation views, in which a tissuesection 16, which has been sliced by a knife 10, is adhered onto a tape32 made of a transparent film; FIG. 4 is a schematic view of a tissuesection manufacturing device of another embodiment; FIG. 5 is anexplanation view explaining the tissue section 16, which has been cutoff from the frozen organism specimen 12 and which curls to outside ofthe slicing surface of the frozen organism specimen 12; and FIGS. 6A and6B are explanation views explaining the conventional method ofcollecting the tissue section 16, in which the curl is manuallyrestricted.

PREFERRED EMBODIMENTS OF THE INVENTION

A tissue section manufacturing device of an embodiment of the presentinvention is shown in FIG. 1. FIG. 1 is a schematic view of the device,which manufactures the tissue section from a frozen organism specimen.

The device shown in FIG. 1 has slicing means 20, which slices a frozenorganism specimen 12 along a slicing surface. The slicing means 20comprises: a holding section 28 including a clamper 26 for holding a jig25, to which the frozen organism specimen 12 is fixed; a servo motor 22for vertically moving the holding section 28 as driving means; and aservo motor 24 for moving the holding section 28 rightward and leftward.Further, a knife 10 for slicing the frozen organism specimen 12 isprovided to a rotatable cylinder 11, and an angle of the knife 10 withrespect to the slicing surface of the frozen organism specimen 12 can beadjusted by the cylinder 11.

Running means 30 makes a tape 32, which is made of a transparent film(hereinafter sometimes referred to “the tape 32”), run apart from theslicing surface of the frozen organism specimen 12. The running means 30is provided to an attachment board 31, which is vertically provided on abase 71, and an attachment plate 46, which is separated from theattachment board 31.

Guide rollers 34 and 34, which support a tape cylinder 36 on which thetape 32 is wound, pressing rollers 42 and 42, which hold the tape 32with a driving roller 44, and a braking roller 33, which is biased by aspring 37 so as to contact the tape cylinder 36 with a prescribed force,are provided to the attachment board 31. Prescribed tension is given tothe tape 32 between the braking roller 33 and the driving roller 44.

Further, a guide roller 35, which guides the tape 32 extended from thetape cylinder 36, a close roller 38, which is located nearest to theslicing surface of the frozen organism specimen 12, and a guide roller40 are provided to the attachment plate 46.

The guide roller 40 adjusts an angle of the tape 32 with respect to theclose roller 38, and a position of the guide roller 40 is adjusted so asto easily adhere tissue sections, which are sliced off from the frozenorganism specimen 12 by the knife 10, to the tape 12.

By employing the running means 30, the tape 32, which has been drawnfrom the tape cylinder 36 with prescribed tension by the driving roller44, is wound by a collecting roller (not shown) via the guide roller 35,the close roller 38, which is located nearest to the slicing surface ofthe frozen organism specimen 12, the guide roller 40 and a space betweenthe pressing rollers 42, 42 and the driving roller 44.

Note that, a preferable diameter of the close roller 38 is 2-40 mm.

The device shown in FIG. 1 has means for adjusting a distance betweenthe tape 32, which is driven by the running means 30, and the frozenorganism specimen 12. The distance adjusting means is constituted by: aplate 56, which is attached to a base member 58 by screws; and anattachment plate 46, which is attached to the plate 56 by screwsinserted in a vertical slit 54.

Further, the device has a screw 48, which moves the plate upward anddownward when the screws inserted in the vertical slit 54 are loosened,and a screw 50, which moves the plates 56 and 46, which are integratedby driving the screws in the slit 54 of the plate 46, rightward andleftward.

When the knife 10 starts to slice the frozen organism specimen 12 thedistance by the distance adjusting means adjusts the slicing surface ofthe frozen organism specimen 12 and the tape 32 so as to make a tip partof the tissue section 16, which has curled to outside of the slicingsurface of the frozen organism specimen 12, contact the tape.

In the device shown in FIG. 1, the close roller 38, which is locatednearest to the slicing surface of the frozen organism specimen 12, ismoved so as to adjust the distance between the tape 32, which is guidedby the close roller 38, and the slicing surface of the frozen organismspecimen 12.

As shown in FIG. 2, a shortest distance C between a slice starting pointP, at which an edge of the knife 10 contacts the slicing surface of thefrozen organism specimen 12 and starts the slice, and the close roller38 by moving the close roller 38.

When the distance C is adjusted, preferably the center O of the closeroller 38 is moved on a bisector line N of an intersection angle αbetween a line M extended from a cutting surface of the knife 10, whichintersects a line R extended from the slicing surface of the organismspecimen 12 at an angle θ, and the line R extended from the slicingsurface of the organism specimen, which intersect at the slice startingpoint P, so as to prevent the close roller 38 from interfering with theknife 10 and the slicing surface of the frozen organism specimen 12.

Note that, the guide roller 40, which is attached to the plate 46 towhich the close roller 38 is attached, is moved together with the closeroller 38 when the distance between the tape 32, which is guided by theclose roller 38, and the slicing surface of the frozen organism specimen12.

Further, the device shown in FIG. 1 has means for adjusting temperaturedifference between the tape 32 and the organism specimen 12 so as toallow the tip part of the tissue section 16 to adhesively abut on oneside of the tape 32.

The temperature adjusting means comprises: a heater 60, which isprovided in a tape running path and which heats the tape 32 toprescribed temperature; a temperature control unit 64, which circulatesair in an insulating box 62, which insulates the slicing means 20, theknife 10, the close roller 38, etc., and maintains temperature of theair at the prescribed temperature; and heat exchanging tubes 65 and 66,which cools the slicing means 20 and the knife 10 so as to maintain aform of the frozen organism specimen 12. The heat exchanging tube 65cools the holding section 28, which includes the clamper 26 for holdingthe jig 25 to which the frozen organism specimen 12 is fixed; the heatexchanging tube 66 cools the knife 10 and a part of the cylinder 11.Further, The temperature control unit 64 has a disinfecting filter 70and a prefilter 72 so as to cool and disinfect the air circulating inthe insulating box 62.

In the temperature adjusting means, the tissue section 16 (see FIG. 5)can be sliced off from the frozen organism specimen 12 in an atmosphere,in which temperature is maintained to maintain the form of the frozenorganism specimen 12, by the temperature control unit 64 and the heatexchanging tubes 65 and 66. To manufacture the good tissue section 16 byslicing the frozen organism specimen 12, preferable temperature of theknife 10 and the frozen organism specimen 12 are −1° C.-−40° C.

On the other hand, when the tape 32 is guided by the close roller 38,the tape is heated by a heater 60 provided in the insulating box 62 soas to adhere the tissue section 16, which contacts the tape. The heater60 may be a known heater, e.g., heat block, cartridge heater, tapeheater.

To adhere the entire tissue section 16 onto the tape 32, whosetemperature has been adjusted, without breaking and forming creases, itis necessary to run the tape 32, on which the tip part of the tissuesection 16 has been adhesively abut, at a speed in synchronism with aslicing speed of the tissue section 16.

Thus, in the device shown in FIG. 1, a control section 21 acts as meansfor synchronizing the running speed of the tape 32 with the slicingspeed of the tissue section 16 is constituted by a servo motor (notshown) for driving the driving roller 44 and a servo motor 22 forvertically moving the holding section 28 and cutting the tissue section16 off from the frozen organism specimen 12 with the knife 10.

In this description, the word “synchronization” means to move the tape32, to which the tip part of the tissue section 16 has been adhesivelyabut, with respect to the slicing speed of the tissue section 16, at therunning speed such that no breaks and no creases are formed in thetissue section. Preferably, the servo motor 22 is controlled to makeratio (Vt/Vs) of the running speed (Vt) of the tape 32 to a runningspeed (Vs) of the holding section 28 1.2-0.8.

Note that, the servo motors assembled in the device shown in FIG. 1 maybe replaced with linear motors, stepping motors, etc.

In the device shown in FIG. 1, when the tissue section 16 is sliced offfrom the frozen organism specimen 12, firstly the frozen organismspecimen 12 is made by directly freezing the specimen in liquid nitrogenor dry ice-acetone or by freezing the specimen, which has been embeddedby an adhesive liquid including an ice crystal-inhibitor andcarboxymethylcellulose (CMC), therein. To fix the frozen organismspecimen 12 to the jig 25, the frozen organism specimen 12 is fixed byan ice crystal-inhibitor.

Carboxymethylcellulose may be mixed with the ice crystal-inhibitor, oran OCT compound, which is commercially sold, may be used.

The jig 25, to which the frozen organism specimen 12 has been fixed, isfixed to the clamper 26 of the holding section 28, then the cylinder 11is turned to adjust the angle θ (see FIG. 2) between the cutting surfaceof the knife 10 and the slicing surface of the frozen organism specimen12.

Next, the close roller 38 is moved by the screws 48 and 50 to adjust thedistance C shown in FIG. 2, which is the shortest distance between theclose roller 38 and the frozen organism specimen 12, so that the tippart of the tissue section 16, which has curled to outside of theslicing surface, can adhesively abut, when the knife 10 starts to slicethe frozen organism specimen 12.

As shown in FIG. 2, when the close roller 38 is moved, the center O ofthe close roller 38 is moved on a bisector line N of the intersectionangle α between the line M extended from the cutting surface of theknife 10, which intersects the line R extended from the slicing surfaceof the organism specimen 12 at the angle θ, and the line R extended fromthe slicing surface of the organism specimen, which intersect at theslice starting point P, so that interference among the close roller, theknife 10 and the slicing surface of the frozen organism specimen 12 canbe avoided.

The tape 32, which has been drawn from the tape cylinder 36, is manuallyengaged with the guide roller 35, the heater 60, the close roller 38,the guide roller 40, the driving roller 44 and the collecting roller(not shown).

Next, the device shown in FIG. 1 is turned on, then the driving roller44 is driven to run the tape 32, the heater 60, which is located betweenthe tape cylinder 36 and the close roller 38 and in the insulating box42, heats the running tape 32 and adjusts the temperature of the tape 32guided by the close roller 38 to prescribed temperature, at which a partof ice fixing the form of the frozen organism specimen 12 is melted toallow the tissue section 16 to adhesively abut on the tape 32, e.g.,0-40° C. (preferably 10-30° C.).

Simultaneously, the temperature control unit 64 cools the entireinsulating box 62, the heat exchanging tube 65 for cooling the frozenorganism specimen 12 and the heat exchanging tube 66 for cooling theknife 10 so as to maintain temperature of an atmosphere of slicing thefrozen organism specimen 12 in a range of −1-40° C.

Further, the servo motor 24 is started to move the holding section 28,which has the clamper 26 for holding the jig 25 to which the frozenorganism specimen 12 has been fixed, rightward or leftward a prescribeddistance, so that thickness of the tissue section 16 sliced off from thefrozen organism specimen 12 is defined. The moving distance of theholding section 28, which is moved rightward and leftward by the servomotor 24, has been previously inputted to the control section 21, whichcontrols the servo motor 24.

Then, the servo motor 22 is started to move the holding section 28upward or downward so as to slice the tissue section 16 off from thefrozen organism specimen 12 by the knife 10. The control section 21synchronizes the servo motor 22 with the servo motor (not shown) fordriving the driving roller 44 so as to synchronize the slicing speed ofthe tissue section 16 with the running speed of the tape 32, so that nocrease are formed in the tissue section 16.

A manner for adhering the tissue section 16, which has been sliced offfrom the frozen organism specimen 12 by the knife 10 with the prescribedthickness, onto the running tape 32 will be explained with reference toFIGS. 3A-3E.

In FIG. 1, the jig 25, to which the frozen organism specimen 12 isfixed, is moved upward and downward, but it is moved rightward andleftward in FIGS. 3A-3E.

Firstly, the jig 25 is moved in a direction of an arrow A so as to movethe frozen organism specimen 12 toward a front tip the fixed knife 10[FIG. 3A]. At that time, the tape 32 is already started to run in themoving direction of the jig 25.

The jig 25 is further moved in the direction of the arrow A, then thefront tip of the knife 10 bites the frozen organism specimen 12 so as tostart to slice the tissue section 16 [FIG. 3B].

The sliced tissue section 16 is guided by a sloped cutting surface ofthe knife 10, and it curls to outside of the slicing surface of thefrozen organism specimen 12, then the tip part thereof contacts the tape32, which is guided by the close roller 38 [FIG. 3C]. Temperature of thetape 32 engaging with the close roller 38 is adjusted, by the heater 60shown in FIG. 1, to the prescribed temperature, at which a part of icefixing the form of the frozen organism specimen 12 is melted to allowthe tissue section 16 to adhesively abut on the tape 32, e.g., 0-40° C.(preferably 10-30° C.), so that the tip part of the tissue section 16,which has contacted the tape 32, is adhered onto the tape 32.

The tissue section 16, whose tip part has been adhered on the tape 32,is moved in the direction of the arrow with the tape 32 and cut off fromthe frozen organism specimen 12 with the movement of the jig 25 in thedirection of the arrow A.

At that time, the control section 21 synchronizes the servo motor (notshown) for driving the driving roller 44, which draws the tape 32, withthe servo motor 33 for moving the jig 25 in the direction of the arrowA, so that the slicing speed of the tissue section 16 can synchronizewith the running speed of the tape 32. With this synchronization, a partof the sliced tissue section 16 is moved together with the tape 32, sothat other sliced parts of the tissue section 16 can be continuouslyadhered onto the tape 32 [FIG. 3D]. Therefore, the entire tissue section16 can be adhered on the tape 32 without forming creases, etc.

When the tissue section 16 is completely cut off from the frozenorganism specimen 12, the tissue section 16, which is wholly adhered onthe tape 32, can be manufactured, then the jig 25 is moved in thereverse direction of the arrow A until reaching the position shown inFIG. 3A [FIG. 3E]. During this action, a clearance is formed between thecutting surface of the knife 10 and the frozen organism specimen 12 soas to prevent an interference of the knife 10 and the frozen organismspecimen 12.

When the device returns to the state shown in FIG. 3A and starts thenext slice, the servo motor 24 is started so as to move the frozenorganism specimen 12 the prescribed distance and define the thickness ofthe tissue 16 sliced off from the frozen organism specimen 12.

By repeating the steps shown in FIGS. 3A-3E, a plurality of the tissuesections 16 can be adhered on the tape 32 in slicing order.

The tape 32, on which the tissue sections 16 are adhered, is drawn bythe driving roller 44 and wound on the collecting roller (not shown).

The tissue sections 16, which are adhered on the tape 32 wound on thecollecting roller, are stained, for microscopic observation, in a statesuch that they are adhered on the long tape 32 or the tape 32 are cut toindividually separate the tissue sections 16.

By using the device shown in FIG. 1, a plurality of the tissue sections16, which are made by continuously slicing the frozen organism specimen12, can be manufactured by an inexperienced worker, thereforemicroscopic samples and samples for optical analysis can be easilymanufactured. Even in clinics employing no experienced workers capableof slicing organism specimens without forming creases, rapidpathological examinations of the frozen organism specimens 12 can beperformed.

Further, the tissue sections 16 are automatically sliced off from thefrozen organism specimen 12 in the insulating box 62, in whichtemperature is maintained and air is circulated, so that there is norisk of infection from the frozen organism specimen 12.

The tissue section 16 can be adhered onto the tape 32 without pressingthe tape 32 onto the frozen organism specimen 12. The disadvantages ofthe conventional methods disclosed in the patent gazettes, such asbreaking tissue cells of the tissue section 16, which is caused bypressing the tape 32 thereonto, and uneven thickness of the tissuesection 16, can be prevented, further the rate-controlling stage ofmanufacturing a microscopic sample can be prevented when the tissuesection 16 is adhered onto the tape 32.

In the actions shown in FIGS. 3A-3E of the device shown in FIG. 1,preferably the servo motor 22 is controlled to make the speed of slicingthe tissue section 16 during the steps shown in FIGS. 3A-3C slower thanthe speed of slicing the tissue section 16 in the step shown in FIG. 3D.By controlling the servo motor 22, degree of curling the tissue section16 toward outside of the slicing surface of the frozen organism specimen12 can be controlled. In this case too, preferably the servo motor fordriving the driving roller 44 is controlled to synchronize the runningspeed of the tape 32 with the speed of slicing the tissue section 16.

In the device shown in FIG. 1, the tape 32, on which the tissue sections16 are adhered, is once wound on the collecting roller, but the tissuesections 16, which have been continuously adhered on the tape 32, may bestained without winding thereon.

The tissue sections 16, which are adhered on the tape 32 and stained,may be adhered on substrates, e.g., slide glass, so as to protect themor use them as microscopic samples.

Further, in the device shown in FIG. 1, the knife 10 and the frozenorganism specimen 12 may be separately cooled so as to maintaintemperature of the knife 10 and the frozen organism specimen 12 between−1° C.-−40° C. In this case, cooling pipes, to which refrigerants aresupplied, or Peltier elements may be individually provided to thecylinder 11, to which the knife 10 is attached, and the jig 25, to whichthe frozen organism specimen 25 is fixed. If the knife 10 and the frozenorganism specimen 12 can be separately cooled to the prescribedtemperature from the atmosphere in which they are provided, controllingtemperature of the insulating box 62, which is performed by the heatexchanging tubes 65 and 66 and the temperature control unit 64, is notrequired.

The device shown in FIG. 1 manufactures the tissue sections 16 from thefrozen organism specimen 12, but the device is capable of manufacturingthe tissue sections 16 by slicing an organism specimen 12, whose form isfixed by using an embedding agent, e.g., paraffin, whose melting pointis higher than room temperature.

If the device shown in FIG. 1 manufactures the tissue sections 16 byslicing the organism specimen 12, whose form is fixed by using anembedding agent, e.g., paraffin, whose melting point is higher than roomtemperature, the atmosphere for slicing the organism specimen 12 neednot be cooled. Therefore, the temperature control unit 64 for coolingair in the insulating box 62 is turned off, and supplying therefrigerants to the heat exchanging tubes 65 and 66 are stopped whilemanufacturing the tissue sections 16 by slicing the embedded organismspecimen 12.

In the device, the heater 60, which is provided in the insulating box62, is used as the temperature adjusting means, which adjusts thetemperature difference between the tape 32 and the organism specimen 12,so as to adhere the tip part of the tissue section 16 onto the tape 32.

When the embedded organism specimen 12 passes the close roller 38, theheater 60 melts a part of the embedding agent, which fixes the form ofthe organism specimen 12, so as to allow the tissue section 16 toadhesively abut on the tape 32; for example, the embedding agent isparaffin whose melting point is 60° C., the tape 32 is heated to 65-70°C.

Note that, since the heater 60 is provided in the insulating box 62, ifinner temperature of the insulating box 62 rises higher than prescribedtemperature, the inner temperature of the insulating box 62 may bemaintained 10-25° C. by the temperature control unit 64.

Another tissue section manufacturing device, which is capable ofmanufacturing the tissue section 16 by slicing the organism specimen 12,which is embedded by an embedding agent whose melting point is higherthan room temperature, e.g., paraffin, is shown in FIG. 4.

In the device shown in FIG. 4, the temperature control unit 64 and theheat exchanging tubes 65 and 66, which are used in the device shown inFIG. 1 so as to circulate air in the insulating box 62 and maintaintemperature therein, are omitted.

Thus, the heater 60 is used, as the means for adjusting the temperaturedifference between the tape 32 and the organism specimen 12, so as toadhere the tip part of the tissue section 16 onto the tape 32.

When the embedded organism specimen 12 passes the close roller 38, theheater 60 melts a part of the embedding agent, which fixes the form ofthe organism specimen 12, so as to allow the tissue section 16 toadhesively abut on the tape 32; for example, the embedding agent isparaffin whose melting point is 60° C., the tape 32 is heated to 65-70°C.

Since the heater is provided in the insulating box 62, if innertemperature of the insulating box 62 rises higher than prescribedtemperature, the inner temperature of the insulating box 62 may bemaintained at prescribed temperature by means for ventilating air in theinsulating box 62 or detaching the insulating box 62.

Note that, in FIG. 4, the structural elements shown in FIG. 1 areassigned the same symbols of FIG. 1, and detail explanation is omitted.

In the above embodiments shown in FIGS. 1-4, the tape 32 is atransparent tape so as to use the tape 32 as a cover glass; if the tape32 is not used as the cover glass, for example, the tissue section 16adhered on the tape 32 is transferred onto a slide glass, the tape maybe made of an opaque film.

In the device shown in FIG. 1, the tape 32 is heated by the heater 60provided in the insulating box 62; if outer temperature of theinsulating box 62 is high and the temperature of the tape 32, whichpasses the close roller 38, can be maintained the prescribed temperatureat which the tissue section 16 is adhered onto the tape 32, the step ofheating the tape 32 by the heater 60 can be omitted.

Further, if the tape 32 is continuously run between the step shown inFIG. 3E and the step shown in FIG. 3A, the tape 32 is wastefullyconsumed, so the servo motor for driving the driving roller 44 may bestopped between the step shown in FIG. 3E and the step shown in FIG. 3Aso as to stop the wasteful consumption of the tape 32.

Note that, the organism specimens, which are treated in the devicesshown in FIGS. 1 and 4, may be not only specimens cut off from animals'bodies, which are used in a field of medical science, but also specimenscut off from plants, which are used in a field of agriculture science.

Experiments

The present invention will be explained in detail with reference toresults of experiments.

Experiment 1

In the device shown in FIG. 1, the tissue sections 16 were made byslicing the frozen organism specimen 12, which was cut off from a piglung, liver or muscle.

The tape 32 used in the device shown in FIG. 1 was the transparent filmtape 32, which was made of cellulose acetate and whose width was 24 mm;the organism specimen 12 was made by the steps of: cutting a piece offfrom a pig lung, liver or muscle; embedding the cut piece incarboxymethylcellulose as an ice crystal-inhibitor, and freezing theembedded piece.

The frozen organism specimen 12 was fixed to the jig 25, on whichcarboxymethylcellulose was applied, then the jig 25, to which the frozenorganism specimen 12 had been fixed, was fixed to the clamper 26 of theholding section 28. The cylinder 11, to which the knife 10 was attached,was turned until the angle θ (see FIG. 2) between the slicing surface ofthe frozen organism specimen 12 fixed to the holding section 28 and thecutting surface of the knife 10 became 22.5 degree.

To cool the frozen organism specimen 12 and the part to which the knife10 was fixed or attached, the refrigerant was applied to the heatexchanging tubes 65 and 66 so as to maintain temperature of the knife 10between −16-−20° C. and temperature of the holding section 28 between−21-−25° C. Further, the temperature control unit 64 circulated air inthe insulating box 62 so as to maintain inner temperature of theinsulating box 62 between −15-−25° C.

The position of the close roller shown in FIG. 1, whose diameter was 10mm, was moved by the screws 48 and 50, which are provided to the plates46 and 56, so as to adjust the distance C shown in FIG. 2 between theclose roller 38 and the frozen organism specimen 12 (see FIG. 2) to 0.4mm.

The tape 32 drawn from the tape cylinder 36 was engaged with the guideroller 35, the heater 60, the close roller 38, the guide roller 40, thedriving roller 44 and the collecting roller (not shown) so as to windthe tape, then the servo motor 24 was driven so as to move the holdingsection 28, which had the clamper 26 for holding the jig 25 to which thefrozen organism specimen 12 was fixed, the prescribed distance rightwardand leftward and defined the thickness of the tissue sections 16, whichwere sliced off from the frozen organism specimen 12, 5 μm, further thedriving roller 44 is rotated to start running the tape 32. The tape 32was heated by the heater 60 provided in the insulating box 62 so as tomaintain the temperature of the running tape 32, which was guided by theclose roller 38, between 0-20° C.

Then, the servo motor 22 was driven so as to move the holding section 28upward and downward, so that the tissue sections having the thickness of5 μm were sliced off from the frozen organism specimen 12 by the knife10. The servo motor 22 was synchronized with the servo motor driving thedriving roller 44 so as to synchronize the running speed of the tape 32with the slicing speed of the tissue sections 16, so that no creaseswere formed in the tissue section 16 adhered on the tape 32.

In this experiment, the speed of slicing the tissue sections 16, whosethickness was 5 μm, was 15 times/minute. During the slice, ratio (Vt/Vs)of the running speed (Vt) of the tape 32 to the moving speed (Vs) of theholding section 28 was adjusted to 0.9. As a result, a plurality of goodtissue sections 16, which had no creases, were serially adhered on thetape 32 and arranged in the longitudinal direction thereof withseparations.

Experiment 2

In this experiment, the speed of slicing the tissue sections 16, whosethickness was 5 μm, was 60 times/minute, but other conditions were thesame as those of the experiment 1; the ratio (Vt/Vs) of the runningspeed (Vt) of the tape 32 to the moving speed (Vs) of the holdingsection 28 was 0.9 as well as the experiment 1. Front ends of sometissue sections 16 were broken, but they were usable as microscopicsamples.

Experiment 3

In this experiment, the distance C shown in FIG. 2 between the closeroller 38 and the frozen organism specimen 12 (see FIG. 2) to 2 mm, butother conditions were the same as those of the experiment 1. Marksexisted in some tissue sections 16, but they were accepted.

Experiment 4

The tissue sections 16 were made in the device shown in FIG. 4 by thesteps of: cutting a piece off from the organism specimen 12, e.g., piglung, liver, muscle; fixing the piece by formalin; dehydrating the fixedpiece by ethanol; penetrating the piece by xylene; and embedding thepiece by paraffin, whose melting point was 60° C.

The tape 32 used in the device shown in FIG. 4 was the transparent filmtape 32, which was made of cellulose acetate and whose width was 24 mm,the organism specimen 12, which was embedded by paraffin whose meltingpoint was 60° C., was fixed to the jig 25, and the jig 25 was fixed tothe clamper 26 of the holding section 28. The cylinder 11, to which theknife 10 was attached, was turned until the angle θ(see FIG. 2) betweenthe slicing surface of the frozen organism specimen 12 fixed to theholding section 28 and the cutting surface of the knife 10 became 22.5degree.

The position of the close roller shown in FIG. 4, whose diameter was 10mm, was moved by the screws 48 and 50, which are provided to the plates46 and 56, so as to adjust the distance C shown in FIG. 2 between theclose roller 38 and the frozen organism specimen 12 (see FIG. 2) to 0.4mm.

The tape 32 drawn from the tape cylinder 36 was engaged with the guideroller 35, the heater 60, the close roller 38, the guide roller 40, thedriving roller 44 and the collecting roller (not shown) so as to windthe tape, then the servo motor 24 was driven so as to move the holdingsection 28, which had the clamper 26 for holding the jig 25 to which theorganism specimen 12 was fixed, the prescribed distance rightward andleftward and defined the thickness of the tissue sections 16, which weresliced off from the organism specimen 12, 5 μm, further the drivingroller 44 is rotated to start running the tape 32. The tape 32 washeated by the heater 60 provided in the insulating box 62 so as tomaintain the temperature of the running tape 32, which was guided by theclose roller 38, between 65-70° C.

Then, the servo motor 22 was driven so as to move the holding section 28upward and downward, so that the tissue sections 16 having the thicknessof 5 μm were sliced off from the organism specimen 12 by the knife 10.The servo motor 22 was synchronized with the servo motor driving thedriving roller 44 so as to synchronize the running speed of the tape 32with the slicing speed of the tissue sections 16, so that no creaseswere formed in the tissue section 16 adhered on the tape 32.

In this experiment, the speed of slicing the tissue sections 16, whosethickness was 5 μm, was 15 times/minute. During the slice, ratio (Vt/Vs)of the running speed (Vt) of the tape 32 to the moving speed (Vs) of theholding section 28 was adjusted to 0.9. As a result, a plurality of goodtissue sections 16, which had no creases, were serially adhered on thetape 32 and arranged in the longitudinal direction thereof withseparations.

Note that, the insulating box 62 was ventilated to maintain innertemperature of the insulating box 62 between 15-25° C.

Effects of the Invention

By the present invention, good tissue sections can be easily and safelymade from an organism specimen by an inexperienced worker. Sincemicroscopic samples can be easily manufactured, pathologicalexaminations can be performed in clinics employing no experiencedworkers so that patients can be properly cured.

Even in clinics in rural and remote areas, e.g., island, in which nopathologic doctors are hired, microscopic images of manufactured samplescan be sent to pathologic doctors via a communication network, e.g.,internet, so that patients can be properly cured.

Further, even inexperienced researchers in other fields, e.g., lifescience, who are not familiar with manufacturing morphologic samples,can easily manufacture observation samples.

1. A method for manufacturing a tissue section, which is used for asample for observation by a microscope, comprises the steps of slicingan organism specimen, whose form has been fixed by freezing or by usingan embedding agent, along a slicing surface, characterized by: adjustinga distance between the slicing surface of the organism specimen and oneside of a film and a temperature difference between the organismspecimen and the film when the slicing of the organism specimen isstarted, thereby a tip part of the tissue section curling to the outsideof the slicing surface of the organism specimen is allowed to adhesivelyabut on one side of the film running apart from the slicing surface ofthe organism specimen; and running the film at a speed in synchronismwith a slicing speed of the tissue section after the tip part of thetissue section is allowed to adhesively abut on the one side of thefilm, thereby the entire tissue section cut off from the organismspecimen can be adhered onto the one side of the film.
 2. The methodaccording to claim 1, wherein the temperature difference between theorganism specimen and the film is adjusted by adjusting temperature ofthe film and/or temperature of an atmosphere, in which the organismspecimen is sliced.
 3. The method according to claim 1, whereintemperature of the film is adjusted to temperature, at which a part ofan ice or the embedding agent for fixing the form of the organismspecimen melts and the tissue section is adhered onto the film.
 4. Themethod according to claim 1, wherein temperature of an atmosphere, inwhich the organism specimen is sliced, is adjusted so as to hold thefixed form of the organism specimen.
 5. The method according to claim 1,wherein a plurality of rollers including a close roller, which isprovided nearest to the slicing surface of the organism specimen, areprovided in a film running path, and the distance between the slicingsurface of the organism specimen and the one side of the film isadjusted by moving the close roller to and away from the slicing surfaceof the organism specimen.
 6. The method according to claim 5, wherein acenter of the close roller is moved on a bisector line of anintersection angle between a line extended from a cutting surface of acutting tool, which intersects a line extended from the slicing surfaceof the organism specimen, and the line extended from the slicing surfaceof the organism specimen.
 7. The method according to claim 1, whereinthe film running speed with respect to the slicing speed of the tissuesection is adjusted so as not to form creases in the tissue section andbreak the tissue section.
 8. The method according to claim 1, whereinratio (Vt/Vs) of the film running speed (Vt) to the slicing speed (Vs)of the tissue section is 1.2-0.8.
 9. The method according to claim 1,wherein the film is a transparent film.
 10. A device for manufacturing atissue section, which is used for a sample for observation by amicroscope, comprising: means for slicing an organism specimen, whoseform has been fixed by freezing or by using an embedding agent, along aslicing surface; and means for running a film apart from the slicingsurface of the organism specimen, characterized by: means for adjustinga distance between the slicing surface of the organism specimen and oneside of the film so as to make a tip part of the curling tissue sectioncontact with the outside of the slicing surface of the organism specimenwhen the slicing of the organism specimen is started; means foradjusting temperature between the organism specimen and the film so asto make the tip part of the tissue section, which has been contacted theoutside of the film, adhesively abut on the film; and means forsynchronizing a slicing speed of the tissue section, whose tip part hasbeen adhered on the one side of the film, and a running speed of thefilm so as to adhere the entire tissue section cut off from the organismspecimen onto the one side of the film.
 11. The device according toclaim 10, wherein said temperature adjusting means adjusts temperatureof the film and/or temperature of an atmosphere, in which the organismspecimen is sliced.
 12. The device according to claim 10, furthercomprising means for adjusting temperature of the film to temperature,at which a part of an ice or the embedding agent for fixing the form ofthe organism specimen melts and the tissue section is adhered onto thefilm, said temperature adjusting means being provided in a film runningpath.
 13. The device according to claim 10, further comprising means foradjusting temperature of an atmosphere, in which the organism specimenis sliced, so as to hold the fixed form of the organism specimen. 14.The device according to claim 10, wherein said distance adjusting meanshas a plurality of rollers, which are provided in a film running pathand which include a close roller provided nearest to the slicing surfaceof the organism specimen, and the close roller is capable of moving toand away from the slicing surface of the organism specimen.
 15. Thedevice according to claim 14, wherein a center of the close roller ismoved on a bisector line of an intersection angle between a lineextended from a cutting surface of a cutting tool, which intersects aline extended from the slicing surface of the organism specimen, and theline extended from the slicing surface of the organism specimen.
 16. Thedevice according to claim 10, wherein said synchronizing means has acontrol section, which controls said running means and said slicingmeans to adjust the film running speed with respect to the slicing speedof the tissue section so as not to form creases in the tissue sectionand break the tissue section.
 17. The device according to claim 16,wherein the control section controls said running means and said slicingmeans so as to make ratio (Vt/Vs) of the film running speed (Vt) to theslicing speed (Vs) of the tissue section 1.2-0.8.
 18. The deviceaccording to claim 10, wherein the the film is a transparent film.